Figure 4.

Quantification of iron in bLf under denaturing conditions. The quantification of iron in the apo- and holo-bLf structures was performed to evaluate the release of this ion in the presence of GnHCl with absorbance measurements at 465 nm. The protein was diluted in standard buffer to a final concentration of 3.5 mg/mL. Fe-NTA solution was added to the samples at 50 μM. Data were obtained from three independent experiments and plotted as mean ± SD. Statistical significance of differences in means compared to respective control conditions without iron, and comparison between holo-bLf under different GnHCl conditions are shown in the figure (∗∗∗, p ≤ 0.001; ∗∗, p ≤ 0.01; ∗, p ≤ 0.05; ns = not significant, p > 0.05).